*6.1.2 Genetic assays*

To detect target site mutations, due to resistance to insecticides such as pyrethroids, carbamates and organophosphates in different *Anopheles* species, PCR assay is most commonly used [72]. It helps in detecting kdr mutations in different species of *Anopheles* due to overuse of pyrethroids/DDT [73, 74]. This assay also detected the resistance in vector population at the metabolic level which involves genes such as cytochrome P450 genes, carboxylesterases and glutathione S-transferases in *Anopheles coluzzii.* Profiling of gene expression was carried out by multiplexing followed by qRT-PCR, the findings from these studies proved it as another effective method for detection of insecticide resistance in malarial vectors. Moreover, study had been carried for the polymorphic genes: P450 genes CYP6Z1, CYP6Z3 and CYP6M7 against pyrethroid resistance in the malaria vector *An. funestus.* The analysis reported the changes in amino acids by QTL, which showed the contribution of these polymorphic genes in the insecticide resistance [75]. Thus, the characterizations and genetic profiling methods can improve the understanding regarding the target site mutations and metabolic resistance in malarial vectors.

### *6.1.3 Proteomics assays*

The major driving factors which contribute to the malaria transmission are the age, method of blood feeding and way of infection spread. These factors could be promising target for detecting the insecticide resistance through proteomics. In *Anopheles* mosquitoes, with the help of artificial neural networks (ANNs) and MALDI-TOF/MS, the effect of insecticide resistance on these factors were described [76]. Proteomics detected the contribution of age as one of significant factor to insecticide resistance by using matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry or capillary high-pressure liquid chromatography with linear ion-trap (LTQ )-Orbitrap XL hybrid mass spectrometer which was further quantified by Western Blot leading to detection of protein biomarkers [77]. Proteomics study also detected metabolic resistance and target sited mutations by NCBInr/Protein BLAST and MS/MS-FTMS in *An. gambiae* in Burkina Faso [78]. Multiple resistance, which is emerging as the big issue in insecticide resistance management has also been detected with the help of proteomics like 2D electrophoresis and MALDI TOF in *An. stephensi* [79]. Thus, proteomics analysis can be a promising tool to tackle various obstacles in insecticide resistance management.

## **6.2 Detection of insecticide resistance in filarial vectors**
